LOCUS       sequence-366157-        4633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  To study dynamics of G-actin using NMR spectroscopy..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4633)
  AUTHORS   Ali R, Zahm JA, Rosen MK
  TITLE     Bound nucleotide can control the dynamic architecture of monomeric 
            actin.
  JOURNAL   Nat Struct Mol Biol. 2022 Apr;29(4):320-328. doi: 
            10.1038/s41594-022-00743-5. Epub 2022 Mar 24.
  PUBMED    35332323
REFERENCE   2  (bases 1 to 4633)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1038/s41594-022-00743-5"; journalName: "Nat Struct Mol Biol"; 
            date: "2022-04"; volume: "29"; issue: "4"; pages: "320-328"
COMMENT     Sequence Label: I282L
FEATURES             Location/Qualifiers
     source          1..4633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             624..653
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             669..686
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     terminator      766..1012
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 terminator
                     /note="transcription terminator for AOX1"
     promoter        1027..1440
                     /gene="S. cerevisiae TEF1"
                     /label=TEF1 promoter
                     /note="promoter for EF-1-alpha"
     promoter        1447..1494
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1513..1887
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     terminator      1953..2200
                     /gene="S. cerevisiae CYC1"
                     /label=CYC1 terminator
                     /note="transcription terminator for CYC1"
     primer_bind     complement(1969..1987)
                     /label=CYC1
                     /note="CYC1 transcription termination signal, reverse 
                     primer"
     rep_origin      complement(2275..2863)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2355..2374)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        2945..3883
                     /gene="Pichia pastoris AOX1"
                     /label=AOX1 promoter
                     /note="inducible promoter, regulated by methanol"
ORIGIN
        1 ttattaccat cggcaacgaa agatttcgat gtcctgaggc cctgttccag ccatcatttt
       61 tgggaatgga ggcatgtgga atccacgaga ctacttacaa ttctttgatg aagtgtgctg
      121 cagctattag aaaggacttg tacgcaaata cggtcctgag tggaggaact acgatgtacc
      181 caggtattgc agatagaatg caaaaggaaa tcacggcctt ggctccatct actatgaaaa
      241 ttaaaatcat cgcaccacct gagcgtaagt attctgtttg gataggcggt tctatattgg
      301 cctctcttag tacattccaa caaatgtgga tctctaaaca ggaatacgat gaatcaggac
      361 caagtatcgt ccaccgtaag tgctttggtg gaagtggagg tagtggtgga tcccttgaag
      421 tattgtttca gggcgcttct agaggaggat ctggaggatc aagtggaggt tctgcctctg
      481 ataagcccga catggctgag atagagaagt ttgataaatc taaattgaaa aaaactgaga
      541 cccaagagaa gaatccattg ccatctaagg aaacgattga acaggaaaag caggctggtg
      601 aaagtgcggc cgccagcttt ctagaacaaa aactcatctc agaagaggat ctgaatagcg
      661 ccgtcgacca tcatcatcat catcattgag tttgtagcct tagacatgac tgttcctcag
      721 ttcaagttgg gcacttacga gaagaccggt cttgctagat tctaatcaag aggatgtcag
      781 aatgccattt gcctgagaga tgcaggcttc atttttgata cttttttatt tgtaacctat
      841 atagtatagg attttttttg tcattttgtt tcttctcgta cgagcttgct cctgatcagc
      901 ctatctcgca gctgatgaat atcttgtggt aggggtttgg gaaaatcatt cgagtttgat
      961 gtttttcttg gtatttccca ctcctcttca gagtacagaa gattaagtga gaccttcgtt
     1021 tgtgcggatc ccccacacac catagcttca aaatgtttct actccttttt tactcttcca
     1081 gattttctcg gactccgcgc atcgccgtac cacttcaaaa cacccaagca cagcatacta
     1141 aatttcccct ctttcttcct ctagggtgtc gttaattacc cgtactaaag gtttggaaaa
     1201 gaaaaaagag accgcctcgt ttctttttct tcgtcgaaaa aggcaataaa aatttttatc
     1261 acgtttcttt ttcttgaaaa tttttttttt tgattttttt ctctttcgat gacctcccat
     1321 tgatatttaa gttaataaac ggtcttcaat ttctcaagtt tcagtttcat ttttcttgtt
     1381 ctattacaac tttttttact tcttgctcat tagaaagaaa gcatagcaat ctaatctaag
     1441 ggcggtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     1501 aggaactaaa ccatggccaa gttgaccagt gccgttccgg tgctcaccgc gcgcgacgtc
     1561 gccggagcgg tcgagttctg gaccgaccgg ctcgggttct cccgggactt cgtggaggac
     1621 gacttcgccg gtgtggtccg ggacgacgtg accctgttca tcagcgcggt ccaggaccag
     1681 gtggtgccgg acaacaccct ggcctgggtg tgggtgcgcg gcctggacga gctgtacgcc
     1741 gagtggtcgg aggtcgtgtc cacgaacttc cgggacgcct ccgggccggc catgaccgag
     1801 atcggcgagc agccgtgggg gcgggagttc gccctgcgcg acccggccgg caactgcgtg
     1861 cacttcgtgg ccgaggagca ggactgacac gtccgacgcg gcccgacggg tccgaggcct
     1921 cggagatccg tccccctttt cctttgtcga tatcatgtaa ttagttatgt cacgcttaca
     1981 ttcacgccct ccccccacat ccgctctaac cgaaaaggaa ggagttagac aacctgaagt
     2041 ctaggtccct atttattttt ttatagttat gttagtatta agaacgttat ttatatttca
     2101 aatttttctt ttttttctgt acagacgcgt gtacgcatgt aacattatac tgaaaacctt
     2161 gcttgagaag gttttgggac gctcgaaggc tttaatttgc aagctggaga ccaacatgtg
     2221 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     2281 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     2341 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     2401 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     2461 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     2521 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     2581 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     2641 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta
     2701 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     2761 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     2821 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     2881 ttctacgggg tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag
     2941 atcagatcta acatccaaag acgaaaggtt gaatgaaacc tttttgccat ccgacatcca
     3001 caggtccatt ctcacacata agtgccaaac gcaacaggag gggatacact agcagcagac
     3061 cgttgcaaac gcaggacctc cactcctctt ctcctcaaca cccacttttg ccatcgaaaa
     3121 accagcccag ttattgggct tgattggagc tcgctcattc caattccttc tattaggcta
     3181 ctaacaccat gactttatta gcctgtctat cctggccccc ctggcgaggt tcatgtttgt
     3241 ttatttccga atgcaacaag ctccgcatta cacccgaaca tcactccaga tgagggcttt
     3301 ctgagtgtgg ggtcaaatag tttcatgttc cccaaatggc ccaaaactga cagtttaaac
     3361 gctgtcttgg aacctaatat gacaaaagcg tgatctcatc caagatgaac taagtttggt
     3421 tcgttgaaat gctaacggcc agttggtcaa aaagaaactt ccaaaagtcg gcataccgtt
     3481 tgtcttgttt ggtattgatt gacgaatgct caaaaataat ctcattaatg cttagcgcag
     3541 tctctctatc gcttctgaac cccggtgcac ctgtgccgaa acgcaaatgg ggaaacaccc
     3601 gctttttgga tgattatgca ttgtctccac attgtatgct tccaagattc tggtgggaat
     3661 actgctgata gcctaacgtt catgatcaaa atttaactgt tctaacccct acttgacagc
     3721 aatatataaa cagaaggaag ctgccctgtc ttaaaccttt ttttttatca tcattattag
     3781 cttactttca taattgcgac tggttccaat tgacaagctt ttgattttaa cgacttttaa
     3841 cgacaacttg agaagatcaa aaaacaacta attattcgaa acgaggaatt catgtgtgat
     3901 gaggaggtag ccgccttggt tgtggataac ggttcaggta tgtgtaaagc aggttttgct
     3961 ggggacgacg cccctagagc tgttttccct tctattgtag gtcgaccccg tcatcaaggc
     4021 gtaatggtcg gcatgggtca aaaagactct tacgtgggag acgaagctca atctaaaaga
     4081 ggaatattga cattgaagta cccaattgag catggtatag ttactaactg ggatgacatg
     4141 gagaaaatct ggcatcacac attctacaac gaattgagag tggcacctga agagcatcca
     4201 gtcttattga cagaggctcc tcttaatcca aaagccaata gagaaaaaat gacccaaata
     4261 atgttcgaga ctttcaacac tccagccatg tatgtcgcca tccaagctgt attgtccttg
     4321 tatgcttcag gcagaaccac gggaatagtt ttagactcag gagatggggt ctctcatact
     4381 gtaccaatat acgaaggtta tgctttacct catgctattt tgcgtctaga tttggctggc
     4441 cgtgacctga ctgattacct tatgaaaatt cttacagaac gagggtactc ctttaccaca
     4501 actgctgagc gtgagattgt cagagatatt aaggaaaaat tgtgttacgt tgcccttgat
     4561 tttgaacagg aaatggctac agctgcatct agtagttcac ttgagaagtc ttacgaattg
     4621 ccagacggtc agg
//
